• Journal of Embryo Transfer
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• v.16 no.2
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• pp.133-138
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• 2001

The aim of this study was to identify the method on extended canine semen exposed to freezing as assessed by motility, survival and acrosomal changes following different freezing ramp rates. Five ejaculates collected by digital manipulation twice weekly from three dogs (Shih-Tzu) were added Tris-Egg Yolk (TE) buffer and divided into 4 aliquots according to formulation of our laboratory. After cooling to 4$^{\circ}C$ by ramp rate of 0.6$^{\circ}C$/min, the samples frozen by ramp rates of 1.6$^{\circ}C$/min to -$25^{\circ}C$, 3$^{\circ}C$/min to -35$^{\circ}C$, 8.9$^{\circ}C$/min to -7$0^{\circ}C$ and 19$^{\circ}C$/min to -11$0^{\circ}C$, respectively, and then stored in L$N_2$for 2days. Each sample was evaluated on their motility, survivability and acrosome integrity at different thawing temperature. The ramp rate of 3$^{\circ}C$/min to -35$^{\circ}C$/h for freezing and thawing temperature of 37$^{\circ}C$ obtained the highest results to improve survivability, motile spermatozoa and intact acrosome appearance than other onditions. In conclusion, we may suggest freezing semen for canine artificial insemination is more efficient with freezing at a ramp rate of -3$^{\circ}C$/min to -35$^{\circ}C$ and thawing with a water bath adjusted to 37$^{\circ}C$.

• Journal of the Korea institute for structural maintenance and inspection
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• v.19 no.4
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• pp.67-74
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• 2015

In this study, effects of micropores on the freezing-thawing resistance of high volume slag concrete are reviewed. Concrete was made with slag which contains the ground granulated blast furnace slag(GGBS) and the pig iron preliminary treatment slag(PS) by replacing 0, 40, 70 %, then compressive strength, freezing-thawing resistance, micropores were reviewed. Also, specified design strength, target air contents were set. Deterioration was induced by using 14-day-age specimen which has low compressive strength for evaluating deterioration by freeze-thawing action. As results of the experiment, despite of specified design strength which has been set similarly and ensured target air contents, the pore size distribution of the concrete showed different results. Micropores in GGBS70 specimen have small amount of water which is likely to freeze because there is small amount of pore volume of 10~100 nm size at 0 cycle which has not been influenced by freezing-thawing. For these reasons, it was confirmed that the freezing-thawing resistance performance of GGBS70 is significantly superior than other specimens because relatively small expansion pressure is generated compared to the other specimens.

• Journal of the Korea Concrete Institute
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• v.23 no.1
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• pp.23-30
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• 2011

In this study, deterioration degrees of concrete were investigated at laboratory under seawater attack and cycling freeze-thaw, which are major durability performance deterioration factors of concrete. Deteriorations of mixed concrete using Portland & blended cement were examined by instrumental analysis of changes in relative dynamic modulus of elasticity and compressive strength. After 520 cycles of freeze-thaw, relative dynamic modulus of elasticity and compressive strength of concrete mixed with normal Portland and LHC over 75% showed relatively low resistance of approximately 44% of those values of SRC. Concrete replaced with 50% fine powder of blast furnace slag showed the most excellent freeze-thaw resistance among the tested blended cement concrete.

• Journal of the Korea institute for structural maintenance and inspection
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• v.14 no.1
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• pp.182-189
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• 2010

The strengthening performance of FRPs(Fiber Reinforced Polymers) is directly affected by the environmental conditions such as freezing-thawing and moisture because FRPs are usually bonded on the concrete surface. It is, therefore, strongly required to evaluate a durability of bond between FRPs and concrete as well as FRP materials itself. The freezing-thawing resistance of FRPs is evaluated in this study with the variables of freezing-thawing conditions, types of FRP and freezing-thawing cycles. From the test results, it is found that tensile strength and pull-off strength of CFRP are not affected by the freezing-thawing. On the other hands, those of GFRP show a little degradation because of continuous water immersion during thawing process. But, cautions are needed on the bond durability between FRPs and concrete in case of continuous water supplying from adjacent to the concrete.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.85-92
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• 1999

This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.

• KSBB Journal
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• v.21 no.2
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• pp.110-117
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• 2006

During routine maintenance, animal cell lines are commonly cryopreserved in growth medium containing serum with 10% DMSO. But, in case of bioprocess under the serum-free conditions, including cultivation of cell lines and producing of pharmaceuticals, the cryopreservation should be executed without serum to prevent a cross-contamination. This experiments were performed to investigate the effects of the serum-free cryopreservation on the CHO cells. To improve the survival rates of the cryopreserved CHO cells in serum-free condition, first, the effects of permeable and non-permeable additives for substitute serum on cell viability were investigated. The combination of 10% DMSO and 0.03 M raffinose in MEM-${\alpha}$ without serum indicated 76% of cell viability. However, it did not reach the survival rates(more than 95%) of the conventional cryopreservation. In the second, to evaluate the cryopreservative ability of the serum-free medium(SFM) we compared viability of the CHO cells cryopreserved in the SFMs(Sigma C5467, C4726, and C1707, JBI SF486 and PF486), the cryoprotectant(Genenmed CAN-1000) and the MEM-${\alpha}$ with serum. All solution contained 10% DMSO. As a result of the comparison, cryopreserved cells in the SFMs showed over 95% of viability and appeared predominant viability better than cryoprotectant CAN-1000. Finally, we assessed the stability of the CHO cells in the long-term cryopreservation(LTC) using SFM. Every three months, the cryopreserved CHO cells were thawed to estimate the cell viability and the recovery rates. Then, real-time RT-PCR analyzed the inserted CHO DHFR gene. All results for the LTC appeared the same stability as the serum containing cryopreservation. In the conclusion, it could be seen that the LTC in the SFM can substitute for serum using methods in the bioprocess proceeded by CHO cells for more than 18 months.

• Journal of Animal Science and Technology
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• v.54 no.4
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• pp.283-290
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• 2012

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. The objective of this study was to investigate the effect of taurine, hypotaurine and trehalose as antioxidants on the function of the freezing-thawed sperm in Korean Jeju Black Bull. The semen was cryopreserved with tris egg yolk extendercontaining 7% glycerol and treated with 20mM taurine, hypotaurine and trehalose. Frozen-thawed sperms were evaluated for sperm motility, viability, membrane integrity, acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender containing 7% glycerol only as control. Frozen-thawed semen evaluation clearlyindicated that the addition of taurine or hypotaurine significantly improved (p<0.05) the motility and viability compared to control spermatozoa. Moreover, in membrane integrity, swollen sperm ratio was significantly increased (p<0.05) in taurine, hypotaurine or trehalose compared to control. In sperm acrosome integrity, F pattern ratio was increased (p<0.05) in hypotaurine among treatments, and AR pattern was significantly lowered (p<0.05) in taurine, hypotaurine and trehalose. In assessed sperm fertilizing ability, taurine, hypotaurine or trehalose significantly improved (p<0.05) the ratio of pronucleus formation and SFI. Finally, compared with the control, addition of taurine, hypotaurine or trehalose as an antioxidant to the freezing extender showed more positive effects on the frozen-thawed spermatozoa. It is concluded that the addition of taurine, hypotaurine, or trehalose to the freezing extender could reduce cryodamage of the Korean Jeju Black Bull spermatozoa.

• Tunnel and Underground Space
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• v.22 no.4
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• pp.276-283
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• 2012

Changes in rock properties due to freezing and thawing cycles ranging from $-20^{\circ}C$ to $10^{\circ}C$ were checked for the typical Korean rocks: granite (weathered), limestone, sandstone, tuff, shale and basalt. The porosity, seismic velocity, shore hardness and specific gravity were measured every 10 cycles for each type of rock up to 40 cycles. The specific gravity was rarely changed. Granite (w), shale and basalt decreased gradually in their shore hardness and seismic velocity values, these values for limestone, sandstone and tuff changed only a very little. The porosity increased in the granite (w), shale and basalt, whereas in the others it did not change. Due to the low tensile strength with high porosity, granite (w), shale and basalt were susceptible to the F-T cycles. A linear regression equation was calculated based on the experiment results according to properties and types of rock. The relationship between the freeze-thaw sensitivity (=initial porosity/initial tensile strength) and the coefficients of the regression equation was examined. With additional experimental data, the coefficients of the regression equation can be estimated using the F-T sensitivity. This makes it possible to predict the properties of rock as affected by freeze-thaw weathering by only measuring the initial properties without knowledge of the regression equation coefficients for each type of rock.

• International Journal of Highway Engineering
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• v.11 no.3
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• pp.51-60
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• 2009

This study investigates the fresh state characteristics, strength, chloride ion penetration resistance and freeze-thaw resistance of the suggested high strength-high durability cement concrete pavement. The required workability and air content could be achieved by using an appropriate admixtures. However its dosage should be carefully determined through field trial batches. Compressive strength increased with the increased cement content and, in particular, high cement volume concrete continuously developed strength up to 90 days. No clear relationship, however, existed between flexural strength and cement content. Chloride penetration resistance seemed as a function of curing age rather than the cement content. Freeze-thaw resistance test was conducted using two different coolants, tap water and 4% NaCl solution. When the tap water was used no severe damage was observed up to 300 cycles regardless the air content. Under 4% NaCl solution, specimens of 326kg/$m^3$ cement content showed severe damage with surface scaling. Based on the experimental investigations herein, it is highly recommended that the cement content be greater than 400kg/$m^3$ for strength-high durability cement concrete pavement structures.

• Magazine of the Korea Concrete Institute
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• v.10 no.4
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• pp.153-161
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• 1998

Concrete structures has been deteriorated by and freezing the thawing due to temperature gap. This study was conducted to evaluate durability of concrete which are increasingly demanded recently. Therefore the research of durability must be executed for application of waste foundry sand concrete real structures. Concrete durability properties incorporating waste foundry sand was performed with the variable of W/C ratio. Sand/Waste foundry sand ratio and Air entrainment-Non air entrainment. Cylinder specimens were made and subjected to freezing and thawing cycle at -18$^{\circ}C$ and 4$^{\circ}C$. Dynamic modulus of elasticity were evaluated as F/T cycle increase. The results show that strength of concrete is increased the W/C ratio decrease, the Sand/Waste foundry sand ratio increases when the concrete contains AE agent and decreasing W/C ratio and AE concrete makes improved resistance of freezing and thawing improved. Especially, resistance of freezing and thawing is improved by Fine aggregate/Waste foundry sand ratio which is 50%, 25%, 0% in a row. Therefore it is turn out the waste foundry sand could be applied to concrete from the experiment.